Application of recombinant TAF3 PHD domain instead of anti-H3K4me3 antibody.

ABSTRACT:

Background

Histone posttranslational modifications (PTMs) represent a focal point of chromatin regulation. The genome-wide and locus-specific distribution and the presence of distinct histone PTMs is most commonly examined with the application of histone PTM-specific antibodies. In spite of their central role in chromatin research, polyclonal antibodies suffer from disadvantages like batch-to-batch variability and insufficient documentation of their quality and specificity.

Results

To mitigate some of the pitfalls of using polyclonal antibodies against H3K4me3, we successfully validated the application of a recombinant TAF3 PHD domain as anti-H3K4me3 affinity reagent in peptide array, western blot and ChIP-like experiments coupled with qPCR and deep sequencing.

Conclusion

SUBMITTER: Kungulovski G 

PROVIDER: S-EPMC4802638 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature